Arteriosclerosis, Thrombosis, and Vascular Biology. 2002;22:1761-1768
Published online before print August 15, 2002, doi: 10.1161/01.ATV.0000034022.11764.EC
Published online before print August 15, 2002, doi: 10.1161/01.ATV.0000034022.11764.EC
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© 2002 American Heart Association, Inc.
Brief Reviews |
Detection of Superoxide in Vascular Tissue
From Universitätsklinik Eppendorf (T.M., A.L.K.), Hamburg Germany; the Vitamin Research Institute (I.B.A.), Moscow, Russia; and the Division of Cardiology (D.G.H.), Emory University School of Medicine, Atlanta, Ga.
Correspondence to Thomas Münzel, Universitätsklinik Eppendorf, Abteilung für Kardiologie, Martinistr. 52, 20246 Hamburg, Germany. E-mail muenzel{at}uke.uni-hamburg.de
During the past decade, it has become apparent that reactive oxygen species play a critical role in the genesis of many vascular diseases. The superoxide anion is among the most important of these, not only because of its rapid reaction with NO but also because it serves as a progenitor for many other reactive oxygen species. Although there are many approaches to detecting and quantifying superoxide in chemical systems, its detection in intact tissues is more difficult. The validity of the most popular and frequently used assay for this purpose, lucigenin-enhanced chemiluminescence, has been recently questioned. It has been suggested that lucigenin itself, especially at high concentrations (>50 µmol/L), may act as a source for superoxide via redox cycling. Lower lucigenin concentrations (5 µmol/L) do not participate in redox cycling to an important extent in intact tissues and, therefore, provide an accurate assessment of the rate of superoxide production in such samples. Other useful assays for superoxide include those using the fluorescent dye dihydroethidine, 2-methyl-6-phenyl-3,7-dihydroimidazo(1,2-
)pyrazin-3-one (CLA), and 2-(p-hydroxybenzyl)-6-(p-hydroxyphenyl) 8-benzylimidazo[1,2-]pyrazin-3-one (coelenterazine). The chemiluminescent compound 5-amino-2,3-dihydroxy-1,4-phthalayineidone (luminol) may also be used to detect various reactive oxygen species and may be made specific for various oxidants, such as hydrogen peroxide, superoxide, and peroxynitrite, by altering the experimental conditions. Although each of these methods may be associated with potential artifacts, the use of 
2 different techniques that yield similar results provides a reliable approach for the study of reactive oxygen species in intact vascular tissues.
Key Words: superoxide • intact tissue • lucigenin • NO synthase III uncoupling
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